*Target (protein/gene name): ecto-ATPase *NCBI Gene # or RefSeq#: 10.2210 *Protein ID (NP or XP #) or Wolbachia#: 3ZX0 *Organism (including strain): Balamuthia mandrillaris
Etiologic Risk Group (see link below): N/A */Disease Information (sort of like the Intro to your Mini Research Write up): Balamuthia infection is a rare and often fatal disease. It was first identified from the brain of a pregnant mandrill baboon in 1986, which died in a zoological park, and was associated with fatal human infection in 1991. It is one of the causes of granulomatous amebic encephalitis, a serious infection of the brain and spinal cord. Balamuthia is thought to enter the body when soil containing Balamuthia comes in contact with skin wounds and cuts, or when dust containing Balamuthia is breathed in. The bacteria can then travel to the brain through the blood stream and cause GAE, which is usually fatal.
Essentiality: Extracellular ATP has profound effects on cellular functions causing plasma membrane depolarization, Ca2+ influx and cell death on various types of cells, with the exception of those that express a high level of ATP-breakdown activity on their surface. Ecto-ATPases are also involved in cellular adhesion, cellular cytotoxicity and protection from cytolytic effects of extracellular ATP. Ecto-ATPase activity is an important factor in regulating the transport of solutes, and may have an impact on fungal nutrition and drug resistance. B. mandrillaris exhibit ecto-ATPase activity.
Druggable Target: ATP synthase ecto-a-subunit targeted in research for breast cancer.
-- Show screenshot of BRENDA enzyme mechanism schematic
Enzyme Assay information (spectrophotometric, coupled assay ?, reagents): -- link to Sigma (or other company) page for assay (see Sigma links below) -- -or link (or citation) to paper that contains assay information -- links to assay reagents (substrates) pages. --- List cost and quantity of substrate reagents, supplier, and catalog #
Structure (PDB or Homology model) -- PDB # or closest PDB entry if using homology model: 10.2210 -- For Homology Model option: ---- Show pairwise alignment of your BLASTP search in NCBI against the PDB ---- Query Coverage: ---- Max % Identities: ---- % Positives ---- Chain used for homology:
Current Inhibitors: Expression Information (has it been expressed in bacterial cells): Purification Method: Image of protein (PyMol with features delineated and shown separately):
*Amino Acid Sequence (paste as text only - not as screenshot or as 'code'):
MAHHHHHHVGTGSNDDDDKSPDPTHNKPLPENVKYGIVLDAGSSHTNLYIYKWPAEKENDTGVVQQLEECQVKGPGISKY
AQKTDEIAAYLAECMKMSTERIPASKQHQTPVYLGATAGMRLLRMESKQSADEVLAAVSRSLKSYPFDFQGAKIITGQEE
GAYGWITINYLLGRFKTPGGSTFGALDLGGSSTQITFVPLNSTLEAPETSLQFRLYGTDYTVYTHSFLCYGKDQALWQKL
AQDIQVSSGGILKDPCFYPGYKKVVNVSELYGTPCTKRFEKKLPFNQFQVQGTGDYEQCHQSILKIFNNSHCPYSQCAFN
GVFLPPLQGSFGAFSAFYFVMDFFKKMANDSVSSQEKMTEITKNFCSKPWEEVKASYPTVKEKYLSEYCFSGTYILSLLL
QGYNFTGTSWDQIHFMGKIKDSNAGWTLGYMLNLTNMIPAEQPLSPPLPHST
Length of protein in amino acids: 452 Molecular Weight: 50539.18 kDa Molar Extinction coefficient of your protein at 280 nm wavelength: 70875 M-1cm-1
results
*CDS Gene Sequence (paste as text only): *GC% Content for gene: *CDS Gene Sequence (codon optimized) - copy from output of Primer Design Protocol (paste as text only): *GC% Content for gene (codon optimized):
*NCBI Gene # or RefSeq#: 10.2210
*Protein ID (NP or XP #) or Wolbachia#: 3ZX0
*Organism (including strain): Balamuthia mandrillaris
Etiologic Risk Group (see link below): N/A
*/Disease Information (sort of like the Intro to your Mini Research Write up): Balamuthia infection is a rare and often fatal disease. It was first identified from the brain of a pregnant mandrill baboon in 1986, which died in a zoological park, and was associated with fatal human infection in 1991. It is one of the causes of granulomatous amebic encephalitis, a serious infection of the brain and spinal cord. Balamuthia is thought to enter the body when soil containing Balamuthia comes in contact with skin wounds and cuts, or when dust containing Balamuthia is breathed in. The bacteria can then travel to the brain through the blood stream and cause GAE, which is usually fatal.
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2493082/
Essentiality: Extracellular ATP has profound effects on cellular functions causing plasma membrane depolarization, Ca2+ influx and cell death on various types of cells, with the exception of those that express a high level of ATP-breakdown activity on their surface. Ecto-ATPases are also involved in cellular adhesion, cellular cytotoxicity and protection from cytolytic effects of extracellular ATP. Ecto-ATPase activity is an important factor in regulating the transport of solutes, and may have an impact on fungal nutrition and drug resistance. B. mandrillaris exhibit ecto-ATPase activity.
Druggable Target: ATP synthase ecto-a-subunit targeted in research for breast cancer.
*EC#: 3.6.1.5
Link to BRENDA EC# page:
https://www.brenda-enzymes.org/enzyme.php?ecno=3.6.1.5
-- Show screenshot of BRENDA enzyme mechanism schematic
Enzyme Assay information (spectrophotometric, coupled assay ?, reagents):
-- link to Sigma (or other company) page for assay (see Sigma links below)
-- -or link (or citation) to paper that contains assay information
-- links to assay reagents (substrates) pages.
--- List cost and quantity of substrate reagents, supplier, and catalog #
Structure (PDB or Homology model)
-- PDB # or closest PDB entry if using homology model: 10.2210
-- For Homology Model option:
---- Show pairwise alignment of your BLASTP search in NCBI against the PDB
---- Query Coverage:
---- Max % Identities:
---- % Positives
---- Chain used for homology:
Current Inhibitors:
Expression Information (has it been expressed in bacterial cells):
Purification Method:
Image of protein (PyMol with features delineated and shown separately):
*Amino Acid Sequence (paste as text only - not as screenshot or as 'code'):
MAHHHHHHVGTGSNDDDDKSPDPTHNKPLPENVKYGIVLDAGSSHTNLYIYKWPAEKENDTGVVQQLEECQVKGPGISKY
AQKTDEIAAYLAECMKMSTERIPASKQHQTPVYLGATAGMRLLRMESKQSADEVLAAVSRSLKSYPFDFQGAKIITGQEE
GAYGWITINYLLGRFKTPGGSTFGALDLGGSSTQITFVPLNSTLEAPETSLQFRLYGTDYTVYTHSFLCYGKDQALWQKL
AQDIQVSSGGILKDPCFYPGYKKVVNVSELYGTPCTKRFEKKLPFNQFQVQGTGDYEQCHQSILKIFNNSHCPYSQCAFN
GVFLPPLQGSFGAFSAFYFVMDFFKKMANDSVSSQEKMTEITKNFCSKPWEEVKASYPTVKEKYLSEYCFSGTYILSLLL
QGYNFTGTSWDQIHFMGKIKDSNAGWTLGYMLNLTNMIPAEQPLSPPLPHST
Length of protein in amino acids: 452
Molecular Weight: 50539.18 kDa
Molar Extinction coefficient of your protein at 280 nm wavelength: 70875 M-1cm-1
*CDS Gene Sequence (paste as text only):
*GC% Content for gene:
*CDS Gene Sequence (codon optimized) - copy from output of Primer Design Protocol (paste as text only):
*GC% Content for gene (codon optimized):